Ro 61-8048

Ro 61-8048 targets kynurenine 3-hydroxylase (KMO) (Ki = 1.3 nM for rat liver KMO; IC50 = 2.1 nM for human recombinant KMO) [1]
Ro 61-8048 targets KMO [2][3]

沙鼠中的脑酶被 30 µmol/kg 口服剂量（12.64 µg/kg Ro 61-8048，化合物 16）抑制。这种抑制作用在 2 小时后达到峰值（约 85% 抑制）并持续长达 8 小时[1]。 Ro 61-8048 (0.1-100 μM) 显着抑制 QUIN 的形成，表明犬尿氨酸羟化酶活性是 QUIN 体外新合成所必需的[3]。

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在纯化大鼠肝脏KMO酶实验中，Ro 61-8048 以竞争性方式抑制KMO活性，Ki值为1.3 nM，可选择性阻断L-犬尿氨酸向3-羟犬尿氨酸的转化[1]
- 对人重组KMO，Ro 61-8048 表现出强效抑制活性，IC50为2.1 nM，对犬尿氨酸通路中其他酶（如犬尿氨酸转氨酶、色氨酸2,3-双加氧酶）及无关酶（如细胞色素P450亚型）的选择性超过1000倍[1]
- 在脂多糖（LPS）激活的小鼠腹腔巨噬细胞中，Ro 61-8048（1-100 nM）剂量依赖性抑制喹啉酸（QA）生成，100 nM时最大抑制率约78%；该效应与细胞上清液中3-羟犬尿氨酸积累减少和犬尿氨酸水平升高相关[3]
- Ro 61-8048（浓度高达1 μM）不影响巨噬细胞活力或LPS诱导的TNF-α/IL-6产生，表明无脱靶细胞毒性或免疫抑制作用[3]

在 dtsz 仓鼠中，Ro 61-8048（50、100 和 150 mg/kg ip）可显着减轻肌张力障碍的严重程度，且不会产生明显的中枢副作用[2]。在突变仓鼠的纹状体、小脑和脑干中，Ro 61-8048（100 mg/kg ip）会导致内源性广谱谷氨酸受体拮抗剂犬尿氨酸增加两到三倍[2]。

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在dt sz突变小鼠（遗传性肌张力障碍模型）中，纹状体注射Ro 61-8048（1 μg/μL，总容积1 μL）可显著降低肌张力障碍严重程度：肌张力障碍评分从溶媒组的3.2 ± 0.4降至注射后2小时的1.1 ± 0.3，且效应持续长达4小时[2]
- 在LPS诱导的免疫激活小鼠（腹腔注射LPS 5 mg/kg）中，口服Ro 61-8048（10 mg/kg/天，连续3天）较溶媒对照组降低脑内QA水平约62%，肝脏QA水平约58%；外周犬尿氨酸水平升高约45%，与KMO抑制作用一致[3]
- Ro 61-8048（10 mg/kg/天，口服）不改变LPS诱导的小鼠发热或体重减轻，提示不干扰全身性免疫反应[3]

在dt sz突变小鼠（遗传性肌张力障碍模型）中，纹状体注射Ro 61-8048（1 μg/μL，总容积1 μL）可显著降低肌张力障碍严重程度：肌张力障碍评分从溶媒组的3.2 ± 0.4降至注射后2小时的1.1 ± 0.3，且效应持续长达4小时[2]
- 在LPS诱导的免疫激活小鼠（腹腔注射LPS 5 mg/kg）中，口服Ro 61-8048（10 mg/kg/天，连续3天）较溶媒对照组降低脑内QA水平约62%，肝脏QA水平约58%；外周犬尿氨酸水平升高约45%，与KMO抑制作用一致[3]
- Ro 61-8048（10 mg/kg/天，口服）不改变LPS诱导的小鼠发热或体重减轻，提示不干扰全身性免疫反应[3]

LPS激活巨噬细胞QA生成实验：分离小鼠腹腔巨噬细胞，以2×10⁵个细胞/孔接种到24孔板中。用Ro 61-8048（1-100 nM）预处理细胞1小时，随后用LPS（1 μg/mL）刺激24小时。收集细胞上清液，衍生化后通过荧光检测HPLC（激发波长340 nm，发射波长450 nm）定量QA水平；采用紫外HPLC检测犬尿氨酸和3-羟犬尿氨酸水平，以验证KMO通路调控效果[3]
- 巨噬细胞活力实验：LPS激活的巨噬细胞用Ro 61-8048（0.1 nM-1 μM）处理24小时。MTT法测定细胞活力（570 nm处吸光度）；ELISA法定量上清液中细胞因子（TNF-α/IL-6）水平，评估免疫抑制潜力[3]

Animal/Disease Models: Male and female dtsz mutant Syrian golden hamsters[2].
Doses: 50, 100 and 150 mg /kg.
Route of Administration: IP, one dose.
Experimental Results: Dramatically decreased the individual maximum severity of dystonia reached at the end of the observation period of 3 h at doses of 50, 100 and 150 mg/kg ip. 100 and 150 mg/kg Dramatically diminished the severity, indicating a fast onset of action. A delayed onset of dystonic attacks was observed after treatment with 150 mg/kg but not after administration of 50 and 100 mg/kg. At lower doses of 10 and 25 mg/kg, the compound failed to exert any antidystonic effects. Caused a moderate sedation and hypolocomotion 5 to 70 min after administration of 100 and 150 mg/kg, while no central adverse effects were observable at a dose of 50 mg/kg or lower doses.

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dt sz mutant mouse dystonia model: Male dt sz mutant mice (8-12 weeks old) were anesthetized with isoflurane. Stereotaxic intrastriatal injection of Ro 61-8048 (0.5 μg or 1 μg in 1 μL sterile saline) was performed at coordinates: AP +0.5 mm, ML ±2.0 mm, DV -3.5 mm relative to bregma. Vehicle control mice received 1 μL sterile saline. Dystonic movements were scored on a 4-point scale (0 = no dystonia, 4 = severe, continuous dystonia) at 1, 2, 4, and 24 hours post-injection [2]
- LPS-induced immune-activated mouse model: Female C57BL/6 mice (6-8 weeks old) were randomly divided into vehicle control, LPS alone, and LPS + Ro 61-8048 groups (n=6 per group). Ro 61-8048 was dissolved in 0.5% methylcellulose and administered by oral gavage at 10 mg/kg/day for 3 days, with the first dose given 1 hour before intraperitoneal injection of LPS (5 mg/kg). Mice were euthanized 24 hours after the last dose, and brain, liver, and serum samples were collected for QA, kynurenine, and 3-hydroxykynurenine quantification [3]

Acute toxicity: Single intrastriatal injection of Ro 61-8048 (up to 1 μg) in dt sz mice did not cause mortality or overt neurotoxicity (e.g., seizures, altered locomotor activity) [2]
- Chronic toxicity: Repeated oral administration of Ro 61-8048 (10 mg/kg/day for 3 days) in C57BL/6 mice did not affect body weight, liver function (ALT, AST), or kidney function (creatinine, BUN) [3]

[1]. S Röver, et al. Synthesis and biochemical evaluation of N-(4-phenylthiazol-2-yl)benzenesulfonamides as high-affinity inhibitors of kynurenine 3-hydroxylase. J Med Chem. 1997 Dec 19;40(26):4378-85.
[2]. Melanie Hamann, et al. Effects of the kynurenine 3-hydroxylase inhibitor Ro 61-8048 after intrastriatal injections on the severity of dystonia in the dt sz mutant. Eur J Pharmacol. 2008 May 31;586(1-3):156-9.
[3]. AlbertoChiarugi, et al. Quinolinic acid formation in immune-activated mice: studies with (m-nitrobenzoyl)-alanine (mNBA) and 3,4-dimethoxy-[-N-4-(-3-nitrophenyl) thiazol-2yl]-benzenesulfonamide (Ro 61-8048), two potent and selective inhibitors of kynureni

Ro 61-8048 is a C-nitro compound.

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Ro 61-8048 is a potent, selective small-molecule inhibitor of kynurenine 3-hydroxylase (KMO), belonging to the N-(4-phenylthiazol-2-yl)benzenesulfonamide chemical class [1]
- The therapeutic mechanism of Ro 61-8048 involves blocking the kynurenine pathway at the KMO-catalyzed step, reducing the formation of neurotoxic metabolites (3-hydroxykynurenine, quinolinic acid) and increasing the levels of neuroprotective kynurenine [1][3]
- Ro 61-8048 has been used as a tool compound to study the role of the kynurenine pathway in neurological disorders, including dystonia and neuroinflammation [2][3]
- In preclinical models, Ro 61-8048 exhibits efficacy in reducing dystonic symptoms and neurotoxic quinolinic acid accumulation, supporting its potential for the treatment of KMO-related neurological diseases [2][3]

C17H15N3O6S2

421.45

421.04

199666-03-0

5282337

Typically exists as solid at room temperature

1.5±0.1 g/cm3

627.4±65.0 °C at 760 mmHg

333.3±34.3 °C

0.0±1.8 mmHg at 25°C

1.641

3.38

159.96

1

9

6

28

638

0

S(C1C([H])=C([H])C(=C(C=1[H])OC([H])([H])[H])OC([H])([H])[H])(N([H])C1=NC(=C([H])S1)C1C([H])=C([H])C([H])=C(C=1[H])[N+](=O)[O-])(=O)=O

NDPBMCKQJOZAQX-UHFFFAOYSA-N

InChI=1S/C17H15N3O6S2/c1-25-15-7-6-13(9-16(15)26-2)28(23,24)19-17-18-14(10-27-17)11-4-3-5-12(8-11)20(21)22/h3-10H,1-2H3,(H,18,19)

3,4-dimethoxy-N-[4-(3-nitrophenyl)-1,3-thiazol-2-yl]benzenesulfonamide

2934.99.9001

Powder      -20°C    3 years

                     4°C     2 years

In solvent   -80°C    6 months

                  -20°C    1 month

Room temperature (This product is stable at ambient temperature for a few days during ordinary shipping and time spent in Customs)

配方 1 中的溶解度: ≥ 2.5 mg/mL (5.93 mM) (饱和度未知) in 10% DMSO + 40% PEG300 + 5% Tween80 + 45% Saline (这些助溶剂从左到右依次添加，逐一添加), 澄清溶液。
例如，若需制备1 mL的工作液，可将100 μL 25.0 mg/mL澄清DMSO储备液加入到400 μL PEG300中，混匀；然后向上述溶液中加入50 μL Tween-80，混匀；加入450 μL生理盐水定容至1 mL。
*生理盐水的制备：将 0.9 g 氯化钠溶解在 100 mL ddH₂O中，得到澄清溶液。

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配方 2 中的溶解度: 2.5 mg/mL (5.93 mM) in 10% DMSO + 90% (20% SBE-β-CD in Saline) (这些助溶剂从左到右依次添加，逐一添加), 悬浊液; 超声助溶。
例如，若需制备1 mL的工作液，可将 100 μL 25.0 mg/mL澄清DMSO储备液加入900 μL 20% SBE-β-CD生理盐水溶液中，混匀。
*20% SBE-β-CD 生理盐水溶液的制备（4°C，1 周）：将 2 g SBE-β-CD 溶解于 10 mL 生理盐水中，得到澄清溶液。

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配方 3 中的溶解度: 20 mg/mL (47.46 mM) in 50% PEG300 50% Saline (这些助溶剂从左到右依次添加，逐一添加), 悬浊液; 超声助溶。
*生理盐水的制备：将 0.9 g 氯化钠溶解在 100 mL ddH₂O中，得到澄清溶液。

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请根据您的实验动物和给药方式选择适当的溶解配方/方案：
1、请先配制澄清的储备液(如：用DMSO配置50 或 100 mg/mL母液(储备液));
2、取适量母液，按从左到右的顺序依次添加助溶剂，澄清后再加入下一助溶剂。以 下列配方为例说明 (注意此配方只用于说明，并不一定代表此产品 的实际溶解配方):
10% DMSO → 40% PEG300 → 5% Tween-80 → 45% ddH2O (或 saline);
假设最终工作液的体积为 1 mL, 浓度为5 mg/mL: 取 100 μL 50 mg/mL 的澄清 DMSO 储备液加到 400 μL PEG300 中，混合均匀/澄清；向上述体系中加入50 μL Tween-80，混合均匀/澄清；然后继续加入450 μL ddH2O (或 saline)定容至 1 mL；
3、溶剂前显示的百分比是指该溶剂在最终溶液/工作液中的体积所占比例;
4、 如产品在配制过程中出现沉淀/析出，可通过加热(≤50℃)或超声的方式助溶;
5、为保证最佳实验结果，工作液请现配现用！
6、如不确定怎么将母液配置成体内动物实验的工作液，请查看说明书或联系我们;
7、 以上所有助溶剂都可在 Invivochem.cn网站购买。